Project description:Bitter pit is the most important physiological disorder affecting apples. In order to ascertain the genetic bases of its incidence in apple fruit, a mapping population of ‘Braeburn’ (susceptible to bitter pit) × ‘Cameo’ (resistant to bitter pit) cultivars was used to map the trait over two growing seasons. RNA-Seq on pools of RNA extracted from fruits of three resistant and three susceptible to bitter pit progenies at post-fertilization and full maturity stages, permitted us to identify a number of candidate genes underlying genetic resistance/susceptibility to bitter pit.
Project description:T2R bitter receptors, encoded by Tas2r genes, are not only critical for bitter taste signal transduction but also important for defense against bacteria and parasites. However, little is known about whether and how Tas2r gene expression are regulated. Here, using single-cell assays for transposase-accessible chromatin with sequencing (scATAC-seq), we found that the chromatin accessibility of Tas2rs was highly cell type specific and lipopolysaccharide (LPS)-induced inflammation increased the accessibility of many Tas2rs. scATAC-seq also revealed substantial chromatin remodeling in immune response genes in taste tissue stem cells, suggesting potential long-term effects. Together, our results suggest an epigenetic mechanism connecting inflammation, Tas2r gene regulation, and altered bitter taste, which may explain heightened bitter taste that can occur with infections and cancer treatments.
Project description:Fruit taste is determined by sugars, acids and in some species, bitter chemicals. Attraction of seed-dispersing organisms in nature and breeding for consumer preferences requires reduced fruit bitterness. A key metabolic shift during ripening prevents tomato fruit bitterness by eliminating α-tomatine, a renowned defence-associated Solanum alkaloid. Here, we combined fine mapping with information from 150 re-sequenced genomes and genotyping a 650 tomato core collection to identify nine bitter-tasting accessions including the ‘high α-tomatine’ Peruvian landraces reported by Rick and colleagues (1994). These ‘bitter’ accessions contain a deletion in GORKY, a nitrate/peptide family (NPF) transporter mediating α-tomatine subcellular localization during fruit ripening. GORKY exports α-tomatine and its derivatives from the vacuole to the cytosol and this facilitates the conversion of the entire α-tomatine pool to non-bitter forms, rendering the fruit palatable. Hence, GORKY activity was a significant innovation in the process of tomato fruit domestication and breeding. The experiment was carried out to further prove that GORKY is localized to tonoplast in ripe fruit.
Project description:We used microarrays to investigate gene expression changes in human colon normal fibroblasts exposed to a bitter orange extract enriched in flavanones (and previously subjected to in vitro gastro-duodenal digestion) to determine possible modulatory beneficial effects induced by these plant-derived compounds on the colon cells. Experiment Overall Design: We used ~90% confluent monolayers of colon CCD-18Co cells exposed to either small doses of pre-digested extract from bitter orange containing a mixture of flavanones (Treated) or equivalent quantities of digestive enzymes and salts (Control). Treatments were done in triplicate.
Project description:An expression profiling was conducted to analyse the effect of the bitter tasting compounds denatonium benzoate (DB) or aloin on the transcriptome of human jejunal crypts compared to DMEM-treated crypts. These two bitter compounds are agonists for the human bitter taste receptor TAS2R43. We took advantage of a deletion polymorphism for TAS2R43, that exists in about 33% of the population to compare the effects of the TAS2R43 agonists in obese subjects that express (TAS2R43(+)) or do not express (TAS2R43(-)) TAS2R43. Primary jejunal crypts from lean (multi-organ donors) or obese (RYGB surgery) subjects were cultured for 24 hours and treated for 4 hours with either DMEM (control) or 1 mM DB or 30 µM aloin. In total 48 mRNA samples were subjected to RNAseq analysis.